CN111732548B

CN111732548B - N2-carbamyl aromatic ring-2-aminopyrimidine derivatives and medical application thereof

Info

Publication number: CN111732548B
Application number: CN202010530851.2A
Authority: CN
Inventors: 刘滔; 李佳; 童乐仙; 周宇波; 董晓武; 王培培; 徐高亚; 李佳楠
Original assignee: Zhejiang University ZJU; Shanghai Institute of Materia Medica of CAS
Current assignee: Zhejiang University ZJU; Shanghai Institute of Materia Medica of CAS
Priority date: 2020-06-11
Filing date: 2020-06-11
Publication date: 2022-06-17
Anticipated expiration: 2040-06-11
Also published as: CN111732548A

Abstract

The present invention provides N²-carbamyl aromatic ring-2-aminopyrimidine derivatives and medical application thereof. N provided by the invention²The-carbamyl aromatic ring-2-aminopyrimidine derivative comprises optical isomers and pharmaceutically acceptable salts thereof. Pharmacodynamic studies show that the compound has FLT3 inhibitory activity, has proliferation inhibitory activity on various leukemia cell strains, has moderate inhibitory action on breast cancer cells, is effective on AML multiple mutations such as internal tandem repeat mutation of a juxtamembrane domain and D835 point mutation of an activation loop in a kinase domain, has almost no inhibitory action on c-KIT, can overcome drug resistance brought by point mutation in clinic, can reduce toxic and side effects of bone marrow inhibition, and can be applied to preparation of antitumor drugs. N is a radical of²Structures of formulae Ia and Ib of carboxamide aromatic ring-2-aminopyrimidine derivatives are as follows:

Description

N2-carbamyl aromatic ring-2-aminopyrimidine derivatives and medical application thereof

Technical Field

The invention relates to the field of medicines, in particular to N²-carbamoyl aromatic ring-2-aminopyrimidine derivatives and their pharmaceutical use, mainly N²-carbamoyl aromatic ring-2-aminopyrimidine derivatives optical isomers, salts or solvates and application thereof in preparing antitumor drugs serving as Fms-like tyrosine kinase 3(FLT3) inhibitors.

Background

Fms-like tyrosine Kinase 3(FLT3), also known in mice as Fetal Liver Kinase 2 (Fetal Liver Kinase-2, FLK2) and Stem Cell Kinase-1 (Stem Cell Kinase-1, STK1), is a type III receptor tyrosine Kinase with Stem Cell growth factor receptor (c-Kit), macrophage colony stimulating factor receptor (Fms), and Platelet Derived Growth Factor Receptor (PDGFR), among others. FLT3 is mainly distributed in hematopoietic cells and neural tissues, and is also present in placenta, gonads, and brain tissues, which play a key role in the proliferation of hematopoietic cells and lymphocytes. The abnormal activation of FLT3 is closely related to the occurrence and development of various tumors, especially Acute Myelogenous Leukemia (AML), and the cure rate of AML patients with high expression of FLT3 is only 30-40% through standard treatment even after bone marrow transplantation. Among them, mutations in FLT3 are the major cause of high relapse rate and poor prognosis in AML patients. At present, the development of inhibitors targeting FLT3 is a hot spot for the study of the treatment of hematological malignancies.

At present, many FLT3 small molecules and biological agents are in clinical research, and the first generation FLT3 inhibitors include sunitinib, sorafenib, midostaurin, lestauatinib and tandutinib, etc., and these inhibitors are characterized by lack of selectivity to FLT 3. The first generation of inhibitors, midostaurin, was approved by the FDA for marketing in 2017. The second generation FLT3 inhibitor has much improved selectivity compared with the first generation, and represents the drugs Giltertinib, Quizartinib and the like. Wherein Giltertinib and Quizaritinib are approved in 2018 and 2019 respectively for treating refractory recurrent mutant acute myeloid leukemia. The main problems faced by FLT3 small molecule inhibitors are drug resistance and toxic side effects. The first generation FLT3 inhibitors were originally developed for other targets and indications, were prone to off-target, which can cause some side effects, and were not effective as monotherapies due to dose-limiting toxicity. These inhibitors generally show better inhibitory activity in the preclinical phase, and are less effective when used in AML patients by the clinical phase I/II. The second generation FLT3 inhibitor is obtained by reasonable drug design, has better selectivity than the first generation, has less second generation FLT3 inhibitors entering clinic at present, and mainly meets the defects of drug resistance, secondary drug resistance and some toxic and side effects caused by mutation, such as myelosuppression, QTc interval prolongation and drug resistance caused by D835 and F691 mutation after drug administration of quizartinib.

Disclosure of Invention

The invention aims to provide N²Carbamoyl aromatic ring-2-aminopyrimidine derivatives, including optical isomers and pharmaceutically acceptable salts thereof, have FLT3 inhibitory activity, have excellent therapeutic effects on tumors, particularly hematological tumors, and are effective on various AML mutations such as internal tandem repeat (ITD) mutation of the juxtamembrane domain and D835 point mutation of the activation ring in The Kinase Domain (TKD), with less bone marrow inhibitory toxic side effects.

In order to achieve the purpose, the invention provides N²-the carboxamide aromatic ring-2-aminopyrimidine derivatives have the general formula Ia and Ib:

and optical isomers thereof, pharmaceutically acceptable salts, wherein:

x is selected from N, CH; q is selected from five to eight membered ring lactams;

R₁is selected from-O (CH)₂)_n R_c、NR_d(CH₂)_nR_c、

Wherein:

n is an integer of 0 to 3; h and k are each independently selected from integers of 1 to 4;

R_cis selected from C_1-4Alkyl radical, C_1-4Alkoxy, unsubstituted or substituted C_5-8Cycloalkanes, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_6-12Spirocycloalkanes, unsubstituted or substituted C_6-12Spiro cycloalkanes containing hetero atoms, unsubstituted or substituted C_6-12Bridged cycloalkanes, unsubstituted or substituted C_6-12The bridged cycloalkane containing heteroatom, wherein the heteroatom is at least one of nitrogen and oxygen, and the substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_fUnsubstituted or substituted C_5-6Aromatic heterocyclic ring of said C_5-6The aromatic heterocyclic ring contains 2-3 hetero atoms selected from oxygen, sulfur and nitrogen, and C_5-6The aromatic heterocyclic substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

m is an integer of 0 to 3;

R_dand R_fEach independently selected from H, deuterium, C_1-6Alkyl radical, C_1-6Hydroxyalkyl radical, C_3-6Cycloalkanes;

R₂selected from the group consisting of-C (O) R_a、-CONR_aR_b、-SO₂NR_aR_b、-NR_aCOR_b、-NR_aSO₂R_b(ii) a Wherein:

R_aand R_bEach independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Hydroxyalkyl, unsubstituted or substituted C_3-8Alicyclic, unsubstituted or substituted C_4-12Aliphatic heterocycles, via C_1-6Alkyl chain-linked unsubstituted or substituted C_3-8Alicyclic ring, through C_1-6Alkyl chain-linked unsubstituted or substituted C_4-12Aliphatic heterocyclic, unsubstituted or substituted C_5-6The aliphatic heterocyclic ring and the aromatic heterocyclic ring contain 1-3 heteroatoms selected from oxygen and nitrogen, and the substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

R₃and R₄Each independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Hydroxyalkyl, halogenated C_1-6Alkoxy radical, C_3-8Cycloalkanes, by C_1-6Alkyl chain linked C_3-8Cycloalkanes, halidesPlain and cyano.

Further, preferred compounds of formula Ia of the present invention have the structure of formula II:

and optical isomers thereof, pharmaceutically acceptable salts, wherein:

R₁is selected from-O (CH)₂)_n R_c、NR_d(CH₂)_nR_c、

Wherein:

R_cis selected from C_1-4Alkyl radical, C_1-4Alkoxy, unsubstituted or substituted C_5-8Cycloalkanes, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkanes, unsubstituted or substituted C_8-18Spiro cycloalkane containing heteroatom(s) of nitrogen or oxygen, wherein said substituent(s) are selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_fUnsubstituted or substituted C_5-6Aromatic heterocyclic ring of said C_5-6The aromatic heterocyclic ring contains 2-3 hetero atoms selected from oxygen, sulfur and nitrogen, and C_5-6The aromatic heterocyclic substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

m is an integer of 0 to 3;

R₂selected from the group consisting of-C (O) R_a、-CONR_aR_b、-SO₂NR_aR_b、-NR_aCOR_b、-NR_aSO₂R_bWherein:

R_aand R_bEach independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Hydroxyalkyl, unsubstituted or substituted C_3-8Alicyclic, unsubstituted or substituted C_4-12Aliphatic heterocycles, via C_1-6Alkyl chain linked unsubstituted or substituted C_3-8Alicyclic ring, through C_1-6Alkyl chain-linked unsubstituted or substituted C_4-12Aliphatic heterocyclic, unsubstituted or substituted C_5-6The aliphatic heterocyclic ring and the aromatic heterocyclic ring contain 1-3 heteroatoms selected from oxygen and nitrogen, and the substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

R₃and R₄Each independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Hydroxyalkyl, halogenated C_1-6Alkoxy radical, C_3-8Cycloalkanes, by C_1-6Alkyl chain linked C_3-8Cycloalkanes, halogens, cyano.

More specifically, preferred compounds of the structure of formula (II) of the present invention are selected from:

morpholinyl (5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) methanone

4-methylpiperazino (5- ((4- ((piperidine-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) methanone

N-phenyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide

5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-pyridin-4-ylpyridinecarboxamide

N-cyclopropyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide

N-4-chlorophenyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide

5- (4- ((piperidin-4-methyl) amino-5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- (4- (methylamino-5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- ((4- (((1r,4r) -4-methylcyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- ((4- ((tetrahydropyran-4-ylmethyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- ((4- (((1r,4r) -4-hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide

5- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-cyclopropylpicolinamide

N- (5- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) cyclopropylcarboxamide

5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide

5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylpyridinamide

5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylpyridinamide

5- ((4- (4-hydroxypiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide

5- ((4- (4-aminopiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide

5- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

5- ((4- ((3-Aminobicyclo [3.2.1] octan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide

N- (5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) pyridin-2-yl) methanesulfonamide

5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylpyridine-2-sulfonamide, and optical isomers and pharmaceutically acceptable salts of the above compounds.

Further, preferred compounds of formula Ia of the present invention have the structure of formula III:

and optical isomers thereof, pharmaceutically acceptable salts, wherein:

R₁is selected from-O (CH)₂)_n R_c、NR_d(CH₂)_nR_c、

Wherein:

R_cis selected from C_1-4Alkyl radical, C_1-4Alkoxy, unsubstituted or substituted C_5-8Cycloalkanes, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkanes, unsubstituted or substituted C_8-18Spiro cycloalkane containing heteroatom(s) of nitrogen or oxygen, wherein said substituent(s) are selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_fUnsubstituted or substituted C_5-6Aromatic heterocyclic ring of said C_5-6The aromatic heterocyclic ring contains 2 to 3 aromatic heterocyclic groups selected fromHetero atoms of oxygen, sulfur, nitrogen, said C_5-6The aromatic heterocyclic substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

m is an integer of 0 to 3;

More specifically, the compounds of the general formula (III) according to the invention are preferably selected from:

4- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (((1r,4r) -4-hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylbenzamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylbenzamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methoxybenzamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylbenzamide

4- ((4- (((1s,4s) -4-hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylbenzamide

4- ((4- (((1s,4s) -4-hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) (piperidin-1-yl) methanone

N- (4- ((4- (((1s,4s) -4-hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) acetamide

N- (4- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) cyclopropylcarboxamide

N- (4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) acetamide

N- (4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) cyclopropylcarboxamide

4- ((4- (4-hydroxypiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (4-aminopiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (((1s,4s) -4-aminocyclohexyl ether) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- ((1-methyl-1H-pyrazol-4-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- ((2-azaspiro [4.5] decan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

4- ((4- ((3-Aminobicyclo [3.2.1] octan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide

N- (4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) methanesulfonamide

4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-toluenesulfonamide including optical isomers and pharmaceutically acceptable salts of the above compounds.

Further, preferred compounds of formula Ib of the present invention have the structure of formula IV:

and optical isomers thereof, pharmaceutically acceptable salts, wherein:

q is selected from five-membered to six-membered cyclic lactams;

R₁is selected from-O (CH)₂)_n R_c、NR_d(CH₂)_nR_c、

Wherein:

R_cis selected from C_1-4Alkyl radical, C_1-4Alkoxy, unsubstituted or substituted C_5-8Cycloalkanes, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkanes, unsubstituted or substituted C_8-18The heteroatom-containing spirocycloalkane, wherein the heteroatom is at least one of nitrogen and oxygen, and the substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_fUnsubstituted or substituted C_5-6Aromatic heterocyclic ring of said C_5-6The aromatic heterocyclic ring contains 2-3 hetero atoms selected from oxygen, sulfur and nitrogen, and C_5-6The aromatic heterocyclic substituent is selected from C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy, halogenated C_1-6Alkoxy, halogen;

m is an integer of 0 to 3;

R_dand R_fEach independently selected from H, deuterium, C_1-6Alkyl radical, C_1-6Hydroxyalkyl radical, C_3-6A cycloalkane;

R₃and R₄Each independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Hydroxyalkyl, halogenated C_1-6Alkoxy radical, C_3-8Cycloalkanes, by C_1-6Alkyl chain linked C_3-8Cycloalkanes, halogens, cyano;

more specifically, preferred compounds of the structure of formula (IV) of the present invention are selected from:

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) isoquinolin-1 (2H) -one

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2- (2-hydroxyethyl) isoquinolin-1 (2H) -one

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-cyclopropanemethylisoquinolin-1 (2H) -one

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-cyclopropaneisoquinolin-1 (2H) -one

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -1-oxoisoquinoline-2 (1H) -methanonitrile

6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-difluoromethylisoquinolin-1 (2H) -one

6- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one

6- ((4- ((2-azaspiro [4.5] decan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one

6- ((4- ((3-Aminobicyclo [3.2.1] octan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one

5- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoindolin-1-one

5- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) isoindolin-1-one, and optical isomers and pharmaceutically acceptable salts of the above compounds.

The invention adopts a method known by a person skilled in the art to prepare the N in the invention²Salts of-carbamoylaromatic ring-2-aminopyrimidines. The salt may be organic acid salt, inorganic acid salt, etc., and the organic acid salt includes citrate, fumarate, oxalate, malate, L-malate, D-malate, lactate, camphorsulfonate, p-toluenesulfonate, methanesulfonate, benzoate, tartrate, L-tartaric acid, D-tartaric acid, oxalic acid, succinic acid, maleic acid, lower fatty acid salt (such as formate, acetate), amino acid salt (such as aspartate), etc.; the inorganic acid salt includes hydrohalic acid salt (such as hydrochloride, sulfate, phosphate, nitrate).

The second purpose of the invention is to provide the compounds of general formulas (I) to (IV), and the application of the optical isomers or the pharmaceutically acceptable salts thereof in preparing antitumor drugs. The compounds of the general formulas (I) to (IV) or pharmaceutically acceptable salts thereof provided by the invention can be used alone and/or together with radiotherapy, other chemotherapeutic drugs and the like in the treatment of FLT 3-mediated diseases (including tumors). The tumor is breast cancer, ovarian cancer, lung cancer, prostate cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, leukemia, lymph cancer, neuroblastoma, glioma, head and neck cancer, thyroid cancer, liver cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, skin cancer, and multiple myeloma. Antineoplastic agents that can be used in combination with the compounds provided herein or pharmaceutically acceptable salts thereof include, but are not limited to, at least one of the following: antimetabolites (e.g., gemcitabine, 5-fluorouracil, hydroxyurea, pemetrexed); alkylating agents (e.g., cisplatin, carboplatin); topoisomerase inhibitors (e.g., irinotecan, doxorubicin); small molecule inhibitors (e.g., MEK inhibitors, PARP inhibitors, Scr inhibitors, mTOR inhibitors, PI3K inhibitors, etc.).

The third object of the present invention is to provide a method for preparing compounds and salts represented by general formulas (I) to (IV), which comprises the following steps:

the method comprises the following steps:

the method comprises the following steps of taking 5-trifluoromethyl-2, 4-dichloropyrimidine as a starting material, sequentially carrying out ammoniation and fatty amine substitution to obtain a 5-trifluoromethyl-pyrimidine-2, 4-diamine intermediate, carrying out coupling reaction with an amide intermediate obtained by condensing 5-bromo-2-pyridinecarboxylic acid or p-bromobenzoic acid and corresponding amine, or p-bromobenzyamine, or 5-bromo-2-cyanopyridine or sulfonamide, and carrying out deprotection and cyano hydrolysis by using ethyl acetate saturated with hydrochloric acid to obtain a target compound.

R₁Is C_5-8Alicyclic rings or C_1-3Alkane or C_5-8C containing 1-3 heterocyclic rings selected from O, N or Boc protection_5-8Containing 1 to 3 heterocyclic rings selected from O, N; r₁' is C_3-8Alicyclic rings or C_1-3Alkane or C_5-8Containing 1 to 3 hetero rings selected from O, N;

R₂is C_1-3Alkane or C_3-8Alicyclic, unsubstituted or substituted C_5-6Aromatic or heteroaromatic ring, unsubstituted or substituted C_4-12An aliphatic heterocycle;

R₂is unsubstituted or substituted amide or sulfonamide.

The second method comprises the following steps:

the amide intermediate obtained by condensing unsubstituted or substituted 2-amino-5-bromopyridine or p-bromoaniline with corresponding acid or the 5-trifluoromethyl-pyrimidine-2, 4-diamine intermediate obtained by the sulfonamide method I is subjected to coupling reaction, and the target compound is obtained by removing a protecting group through trifluoroacetic acid.

R₁Is C_5-8Alicyclic rings or C_1-3Alkane or C_5-8C containing 1-3 heterocyclic rings selected from O, N or protected by Boc_5-8Containing 1 to 3 hetero rings selected from O, N;

R₁' is C_3-8Alicyclic rings or C_1-3Alkane or C_5-8Containing 1 to 3 hetero rings selected from O, N;

R₂is C_1-3Alkane or C_3-8An alicyclic ring;

R₂' is unsubstituted or substituted amide, sulfonamide;

R₃is a or C_1-3Alkane or C_1-3An alkoxy hydrocarbon.

The third method comprises the following steps:

and (3) carrying out coupling reaction on unsubstituted or substituted 4-bromoxynil and the 5-trifluoromethyl-pyrimidine-2, 4-diamine intermediate prepared by the first method, hydrolyzing a cyano group by 75% of concentrated sulfuric acid, and removing a protecting group to obtain the target compound.

R₁' is C_3-8Alicyclic rings or C_1-3Alkane or C_5-8Containing 1 to 3 oxygen atoms selected from O,A heterocycle of N;

R₃is a or C_1-3Alkane or C_1-3An alkoxy hydrocarbon.

The method four comprises the following steps:

coupling reaction is carried out on 6-bromo-2H-isoquinoline-1-ketone or 6-bromo-2H-isoquinoline-1-ketone and the 5-trifluoromethyl-pyrimidine-2, 4-diamine intermediate prepared by the first method through halogenated hydrocarbon or halogenated alcohol substitution products, and finally, a protecting group is removed to obtain the target compound.

R₂is C_1-6Alkyl or C_3-8Alicyclic or halogenated C_1-6Alkyl or C_1-6Hydroxyalkyl radical through C_1-6Alkyl chain linked C_3-8Cycloalkane or cyano.

The method five comprises the following steps:

dissolving 1.2 equivalents of inorganic acid or organic acid in ethanol solution, slowly dripping into ethanol solution of the compound, adding appropriate amount of diethyl ether solution, vacuum filtering to obtain salt-forming compound, washing with diethyl ether, and drying to obtain corresponding compound salt form.

The compounds and salts of the present invention can be prepared by the above-mentioned methods or similar methods, and the corresponding raw materials are selected according to the difference of substituents and the position of substituents.

The invention provides N²The primary drug effect shows that the compound has FLT3 inhibitory activity, has proliferation inhibitory activity on various leukemia cell strains, has moderate inhibitory action on breast cancer cells, and has various AML inhibitory effectsThe mutation is effective as internal mutation of membrane-proximal domain (ITD) and D835 point mutation of activation ring in kinase domain (TKD), has almost no inhibitory effect on c-KIT, can overcome drug resistance caused by point mutation in clinic, and can reduce toxic and side effects of bone marrow inhibition.

Detailed Description

The present invention is further illustrated by reference to the following examples, which are intended to be illustrative of the invention and are not to be construed as limiting in any way.

Preparation example 1 preparation of Compounds 1 to 7

Step 1.4-chloro-5-trifluoromethyl-2-aminopyrimidine (1-2)

Dissolving 2, 4-dichloro-5-trifluoromethylpyrimidine (6.5g,30.0mmol) in acetonitrile (30mL), slowly adding ammonia water (20mL) dropwise under ice bath, moving to room temperature for reaction for 40min after dropping, and recovering the solvent by reduced pressure distillation to obtain a residue. Purifying with silica gel column chromatography, and eluting with PE: EA (6:1) to obtain white solid intermediate 1-2. Yield: 46 percent;¹H NMR(500MHz,CDCl₃) δ8.47(s,Ar-H,1H),5.57(s,NH,2H)；ESI-MS:m/z＝217[M+H]⁺。

step 2.1-Boc-4- (((2-amino-5- (trifluoromethyl) pyrimidin-4-yl) amino) methyl) piperidine (1-3)

Dissolving the intermediate 1-2(593mg,3.0mmol) in anhydrous methanol (12mL), sequentially adding triethylamine (303mg,3.0 mmol) and 1-Boc-4-aminomethylpiperidine (771mg,3.6mmol), heating and refluxing for 8h, and recovering the solvent by reduced pressure distillation to obtain a residue. Purifying by silica gel column chromatography with PE, EA (2:1) is used as eluent to obtain 1-3 white solid. Yield: 80 percent;¹H NMR(500MHz,DMSO-d₆)δ7.95(s,Ar-H,1H),6.71(m,NH,1H),6.70(s,NH₂,2H),4.02–3.79 (m,CH₂,2H),3.26(t,J＝6.5Hz,CH₂,2H),2.81–2.55(m,CH₂,2H),1.82(tdq,J＝11.0,7.5,3.5 Hz,CH,1H),1.57(dd,J＝13.0,4.0Hz,CH₂,2H),1.38(s,CH₃×3,9H),0.99(qd,J＝12.5,4.0Hz, 2H)；ESI-MS:m/z＝376[M+H]⁺。

step 3, synthesizing the intermediate 1-5-1-11

The compound 5-bromo-2-pyridinecarboxylic acid (404mg,2.0mmol) was dissolved in dichloromethane-free (10mL), and the corresponding amine (2.2mmol), HOBT (324mg,2.4mmol), EDCI (575mg,3.0mmol), DIPEA (383mg,3.0mmol, 522. mu.L) were added under stirring in an ice bath, followed by transfer to room temperature for 3-5 h. After the reaction is finished, the solvent is removed by reduced pressure distillation, and the mixture is purified by silica gel column chromatography, and PE: EA (5: 1-1: 1) is used as an eluent to obtain white or light yellow solid 1-5-1-11. Yield: 60-80%.

3.15-bromo-2-morpholinocarbonylpyridine (1-5)¹H NMR(500MHz,DMSO-d₆)δ8.73(d,J＝2.5Hz, Ar-H,1H),8.20(dd,J＝8.5,2.5Hz,Ar-H,1H),7.59(d,J＝8.5Hz,Ar-H,1H),3.68–3.62(m, CH₂,4H),3.57–3.52(m,CH₂,2H),3.43–3.41(m,CH₂,2H).ESI-MS:m/z＝271[M+H]⁺。

3.25-bromo-2-piperazinecarbonylpyridine (1-6)¹H NMR(500MHz,DMSO-d₆)δ8.72(d,J＝2.5Hz, NH,1H),8.18(dd,J＝8.5,2.5Hz,Ar-H,1H),7.55(d,J＝8.5Hz,Ar-H,1H),3.62(dd,J＝6.5,4.0 Hz,CH₂,2H),3.37(d,J＝5.0Hz,CH₂,2H),2.37(t,J＝5.0Hz,CH₂,2H),2.27(t,J＝5.0Hz,CH₂, 2H),2.19(s,CH₃,3H).ESI-MS:m/z＝284[M+H]⁺。

3.35-bromo-2-N-phenylpyridinamides (1-7)¹H NMR(500MHz,DMSO)δ10.64(s,NH,1H), 8.88–8.83(m,Ar-H,1H),8.32(dd,J＝8.5,2.5Hz,Ar-H,1H),8.09(dd,J＝8.5,0.5Hz,Ar-H, 1H),7.89(dd,J＝8.5,1.0Hz,Ar-H,2H),7.39–7.33(m,Ar-H,2H),7.15–7.10(m,Ar-H,1H). ESI-MS:m/z＝277[M+H]⁺。

45-bromo-2-N-pyridin-4-ylpyridinamide (1-8) H NMR (500MHz, DMSO) δ 11.04(s, NH, 1H), 8.91-8.84 (M, Ar-H,1H),8.49(dd, J ═ 5.0,1.5Hz, Ar-H,2H),8.34(dd, J ═ 8.5,2.5Hz, Ar-H,1H),8.10(dd, J ═ 8.5,0.5Hz, Ar-H,1H), 7.97-7.90 (M, Ar-H,2H) ESI-MS: M/z ═ 278 [ M + H,2H ], [ M + H ]]⁺。

3.55-bromo-N-cyclopropylpicolinamide (1-9)¹H NMR(500MHz,CDCl₃)δ8.56(dd,J＝2.5,1.0 Hz,Ar-H,1H),8.09(dd,J＝8.5,1.0Hz,Ar-H,1H),7.97(dd,J＝8.5,2.5Hz,Ar-H,1H),7.94– 7.87(m,NH,1H),2.92(tq,J＝7.5,4.0Hz,H,1H),0.91–0.85(m,CH₂,2H),0.68–0.62(m,CH₂, 2H).ESI-MS:m/z＝241[M+H]⁺。

3.65-bromo-N-4-chlorophenyl picolinamide (1-10)¹H NMR(500MHz,DMSO-d₆)δ10.82(s,NH, 1H),8.85(d,J＝2.0Hz,Ar-H,1H),8.32(dd,J＝8.5,2.0Hz,Ar-H,1H),8.08(d,J＝8.5Hz,Ar-H, 1H),7.98–7.90(m,Ar-H,2H),7.45–7.36(m,Ar-H,2H).ESI-MS:m/z＝311[M+H]⁺。

3.7 (5-Bromopyridin-2-yl) (4- (4-methylpiperazin-1-yl) piperidin-1-yl) methanone (1-11)¹H NMR(500MHz, CDCl₃)δ8.63(dd,J＝2.5,1.0Hz,Ar-H,1H),7.91(dd,J＝8.5,2.5Hz,Ar-H,1H),7.52(dd,J＝8.5,1.0Hz,Ar-H,1H),4.79–4.65(m,CH₂-a,1H),4.11–3.91(m,CH₂-b,1H),3.09–3.03(m, CH₂-a,1H),2.84–2.78(m,CH₂-b,1H),2.61(s,CH₂×2,4H),2.52(m,CH₂×2,CH,5H),2.29(s, CH₃,3H),1.97–1.93(m,CH₂-a,1H),1.82–1.78(m,CH₂-b,1H),1.56(m,CH₂,2H).ESI-MS: m/z＝367[M+H]⁺。

Step 4, synthesizing compounds 1-7

Under the protection of nitrogen, anhydrous dioxane (3mL) was added to a mixture of compound 1-3(113mg,0.3mmol), one of intermediates (0.36mmol) in 1-5 to 1-11, tris (dibenzylideneacetone) dipalladium (14mg,0.015mmol), 4, 5-bis diphenylphosphine-9, 9-dimethylxanthene (22mg,0.0375 mmol), and cesium carbonate (195mg,0.6mmol), and the mixture was stirred under reflux overnight. Suction filtering, and recovering solvent under reduced pressure to obtain residue. Purifying by silica gel column chromatography with CH₂Cl₂EtOH (25:1) as eluent, 2mL CH was used after intermediate was obtained₂Cl₂Dissolving, adding 1mL of CF under ice-bath condition₃COOH, at room temperature for 40 min. Recovering solvent under reduced pressure to obtain residue, dissolving with anhydrous methanol (2mL), adjusting pH to more than 8 with 1N NaOH aqueous solution, recovering solvent under reduced pressure to obtain residue, purifying with silica gel column chromatography, and purifying with CH₂Cl₂MeOH, TEA (30:1: 1-20: 1:1) as eluent to give a white solid. Yield: 35-55 percent.

4.1 morpholinyl (5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) methanone (Compound 1)¹H NMR(500MHz,DMSO-d₆)δ10.02(s,NH 1H),8.93(d,J＝2.5Hz,Ar-H,1H),8.27 (dd,J＝8.5,2.5Hz,Ar-H,1H),8.24(s,Ar-H,1H),7.61(d,J＝8.5Hz,Ar-H,1H),7.37(m,Ar-H, 1H),3.69–3.57(m,CH₂,8H),3.35(t,J＝6.5Hz,CH₂,2H),3.14(dt,J＝12.5,3.5Hz,CH₂,2H), 2.66(td,J＝12.5,2.5Hz,CH₂,2H),1.99–1.92(m,CH,1H),1.74–1.71(m,CH₂,2H),1.35–1.27 (m,CH₂,2H).ESI-MS：m/z＝466[M+1]⁺。

4.24-Methylpiperazinyl (5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) methanone (Compound 2)¹H NMR(500MHz,DMSO-d₆)δ10.03(s,NH,1H),8.95(d,J＝2.5Hz,Ar-H, 1H),8.24(d,J＝2.5Hz,Ar-H,1H),8.23(s,Ar-H,1H),7.55(d,J＝8.5Hz,Ar-H,1H),7.39(s,NH, 1H),3.53(d,J＝10.0Hz,CH₂,2H),3.31(t,J＝6.5Hz,CH₂,2H),2.96(d,J＝12.0Hz,CH₂,2H), 2.42(t,J＝12.0Hz,CH₂,2H),2.35(t,J＝5.0Hz,CH₂,2H),2.28(t,J＝5.0Hz,CH₂,2H),2.18(s, CH₃,3H),1.63-1.60(m,CH₂,2H),1.19–1.03(m,CH₂,2H).ESI-MS：m/z＝479[M+1]⁺。

4.3N-phenyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide (Compound 3)¹H NMR(500MHz,DMSO-d₆)δ10.43(s,NH,1H),10.19(br,NH,1H),8.99(d,J＝2.5Hz, Ar-H,1H),8.54(dd,J＝8.5,2.5Hz,Ar-H,1H),8.27(s,Ar-H,1H),8.07(d,J＝8.5Hz,Ar-H,1H), 7.90(d,J＝1.0Hz,Ar-H,1H),7.88(d,J＝1.0Hz,Ar-H,1H),7.40-7.39(m,Ar-H,1H),7.38– 7.32(m,Ar-H,2H),7.16–7.03(m,NH,1H),3.36(d,J＝6.5Hz,CH₂,2H),2.94(dt,J＝12.5,3.5 Hz,CH₂,2H),2.45–2.33(m,CH₂,2H),1.87-1.80(m,CH,1H),1.66–1.56(m,CH₂,2H),1.14– 1.06(m,CH₂,2H)；ESI-MS：m/z＝472[M+1]⁺。

4.45- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-pyridin-4-ylpyridinecarboxamide (Compound 4)¹H NMR(500MHz,DMSO-d₆)δ10.83(s,NH,1H),10.24(s,NH,1H),9.00(d,J＝ 2.5Hz,Ar-H,1H),8.56(dd,J＝8.5,2.5Hz,Ar-H,1H),8.47(d,J＝5.5Hz,Ar-H,2H),8.27(s, Ar-H,1H),8.08(d,J＝8.5Hz,Ar-H,1H),7.92(d,J＝5.5Hz,Ar-H,2H),7.41(s,NH,1H),3.35(t, J＝6.5Hz,CH₂,2H),2.96(d,J＝12.0Hz,CH₂,2H),2.47–2.36(m,CH₂,2H),1.87–1.81(m,CH, 1H),1.70–1.55(m,CH₂,2H),1.15-1.07(m,CH₂,2H)；ESI-MS：m/z＝473[M+1]⁺。

4.5N-cyclopropyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide (Compound 5)¹H NMR(500MHz,DMSO-d₆)δ10.07(br,NH,1H),8.84(s,Ar-H,1H),8.50(d,J＝5.0 Hz,NH,1H),8.44(d,J＝8.5Hz,Ar-H,1H),8.23(s,Ar-H,1H),7.91(d,J＝8.5Hz,Ar-H,1H), 7.34(s,NH,1H),2.95–2.87(m,CH₂,2H),2.86–2.84(m,CH,1H),2.34(t,J＝12.0Hz,CH₂,2H), 1.86–1.72(m,CH,1H),1.58–1.56(m,CH₂,2H),1.32–1.16(m,CH₂-a,1H),1.09–0.99(m, CH₂,2H),0.82(m,CH₂-b,1H),0.72–0.59(m,CH₂×2,4H)。ESI-MS：m/z＝436[M+1]⁺。

4.6N-4-chlorophenyl-5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridinecarboxamide (Compound 6)¹H NMR(500MHz,DMSO-d₆)δ10.60(br,NH,1H),10.23(s,NH,1H),8.98(d,J＝ 2.5Hz,Ar-H,1H),8.51(dd,J＝8.5,2.5Hz,Ar-H,1H),8.29(s,Ar-H,1H),8.11(d,J＝8.5Hz, Ar-H,1H),7.97–7.91(m,Ar-H×2,2H),7.48(t,J＝6.0Hz,NH,1H),7.43–7.35(m,Ar-H×2,2H), 3.44–3.37(m,CH₂,2H),3.30–3.26(m,CH₂,2H),2.86–2.80(m,CH₂,2H),2.06–2.01(m,CH, 1H),1.87–1.76(m,CH₂,2H),1.46–1.31(m,CH₂,2H).ESI-MS：m/z＝506[M+1]⁺。

4.7(4- (4-methylpiperazin-1-yl) piperidin-1-yl) (5- ((4- ((piperidin-4-methyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) methanone (Compound 7)¹H NMR(500MHz,DMSO-d₆)δ10.06(s,NH,1H),8.89(dd,J ＝2.5,1.0Hz,Ar-H,1H),8.29–8.26(m,Ar-H,1H),8.25(d,J＝1.0Hz,Ar-H,1H),7.57(d,J＝8.5 Hz,Ar-H,1H),7.48(s,NH,1H),4.51–4.49(m,CH₂-a,1H),3.98(s,CH₂-b,1H),3.35(s,CH₂×2, 4H),3.27(d,CH₂×2,4H),2.97–2.92(m,CH₂-a,1H),2.81–2.79(s,CH₂+CH×2,4H),2.76–2.73 (s,CH₂,2H),2.09–1.98(m,CH,1H),1.81–1.78(m,CH₂×2,4H),1.41–1.34(m,CH₂×2,4H), 1.15–1.11(m,CH₂,2H).ESI-MS：m/z＝562[M+1]⁺。

Preparation example 2 preparation of Compounds 8 to 16

Step 1, synthesizing a compound 1-12-1-19

Synthesis procedure with reference to step 2 of example 1, compounds 1-12 to 1-19 were prepared using the corresponding amine instead of 1-Boc-4-aminomethylpiperidine.

1.1N⁴- (tetrahydropyran-4-ylmethyl) -5-trifluoromethylpyrimidine-2, 4-diamine (1-12)¹H NMR(500MHz, CDCl₃)δ8.05(s,Ar-H,1H),5.35–4.99(m,s,NH×3,3H),4.00–3.97(m,CH₂,2H),3.42–3.34 (m,CH₂×2,4H),1.91–1.82(m,CH,1H),1.65–1.60(m,CH₂,2H),1.38–1.30(m,CH₂,2H)； ESI-MS：m/z＝277[M+1]⁺。

1.2 1-Boc-N⁴- (1r,4r) -4-aminocyclohexyl-5-trifluoromethylpyrimidine-2, 4-diamine (1-13)¹H NMR(500 MHz,DMSO-d₆)δ7.96(s,Ar-H,1H),6.76(d,J＝8.5Hz,NH₂,NH₂-a,3H),6.05(d,J＝8.5Hz, NH₂-b,1H),4.19–3.94(m,CH,1H),3.22–3.15(m,CH,1H),1.78–1.73(m,CH₂×2,4H),1.55– 1.42(m,CH₂,2H),1.37(s,CH₃×3,9H),1.24–1.16(m,CH₂,2H)；ESI-MS：m/z＝376[M+1]⁺。

1.3 1-Boc-N⁴- (1s,4s) -4-aminocyclohexyl-5-trifluoromethylpyrimidine-2, 4-diamine (1-14)¹H NMR(500 MHz,DMSO-d₆)δ8.00(s,Ar-H,1H),6.80(br,NH₂,2H),6.59(d,J＝8.0Hz,NH₂-a,1H),5.53(d, J＝8.0Hz,NH₂-b,1H),4.16(qt,J＝8.5,3.5Hz,CH,1H),3.62–3.48(m,CH,1H),1.76–1.69(m, CH₂,2H),1.60–1.51(m,CH₂×3,6H),1.39(s,CH₃×3,9H)；ESI-MS：m/z＝376[M+1]⁺。

1.4N⁴- (1r,4r) -4-aminocyclohexyl-5-trifluoromethylpyrimidine-2, 4-diamine (1-15)¹H NMR(500MHz,CDCl₃) δ8.02(q,J＝1.0Hz,Ar-H,1H),5.16(s,NH₂,2H),4.82(d,J＝7.5Hz,NH,1H),3.96–3.90(m, CH,1H),2.11–1.98(m,CH₂,2H),1.81–1.67(m,CH₂,2H),1.41–1.32(m,CH,1H),1.23–1.12 (m,CH₂,2H),1.11–1.03(m,CH₂,2H),0.91(d,J＝6.5Hz,CH₃,3H)；ESI-MS：m/z＝275[M+1]⁺。

1.5(1r,4r) -4- (2-amino-5-trifluoromethylpyrimidin-4-yl) amino) cyclohexanol (1-16)¹H NMR(500MHz, DMSO-d₆)δ7.96(s,Ar-H,1H),6.73(s,NH₂，2H),5.93(d,J＝8.0Hz,OH,1H),4.57–4.56(m, CH,1H),4.09–4.01(m,CH,1H),1.87–1.79(m,CH₂,2H),1.77–1.73(m,CH₂,2H),1.53–1.38 (m,CH₂,2H),1.25–1.15(m,CH₂,2H)；ESI-MS：m/z＝277[M+1]⁺。

1.61- (2-amino-5-trifluoromethylpyrimidin-4-yl) piperidin-4-ol (1-17)¹H NMR(500MHz,DMSO-d₆) δ8.17(s,Ar-H,1H),6.88(s,NH×2,2H),4.75(d,J＝4.5Hz,OH,1H),3.79–3.75(m,CH₂,2H), 3.73–3.67(m,CH,1H),3.15–3.10(m,CH₂,2H),1.81–1.75(m,CH₂,2H),1.42–1.35(m,CH₂, 2H)；ESI-MS：m/z＝263[M+1]⁺。

1.74-Boc- (1- (2-amino-5-trifluoromethylpyridin-4-yl) piperidine (1-18)¹H NMR(500MHz,DMSO-d₆) δ8.17(s,Ar-H,1H),6.96–6.86(m,NH×3,3H),3.98–3.88(m,CH₂,2H),3.51–3.49(m,CH, 1H),3.02–2.91(m,CH₂,2H),1.79–1.75(m,CH₂,2H),1.37(s,CH₃×3+CH₂,11H)；ESI-MS：m/z ＝362[M+1]⁺。

1.8N⁴-methyl-5-trifluoromethylpyrimidine-2, 4-diamine (1-19)¹H NMR(500MHz,DMSO-d₆)δ7.95(s, Ar-H,1H),6.75–6.74(m,NH×3,3H),2.82(d,J＝4.5Hz,CH₃,3H)；ESI-MS：m/z＝193[M+1]⁺。

Step 2, synthesizing compounds 8-16

Synthesis procedure referring to step 4 of preparation example 1, compounds 8 to 16 were prepared by substituting 1-12 to 1-19 for 1-3 and 1-20 for 1-5 to 1-11. Yield: 35-55 percent.

2.15- (4- ((piperidin-4-methyl) amino-5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (compound 8) ESI-MS: 396[ M + 1] M/z]⁺。

2.25- ((4- ((tetrahydropyran-4-ylmethyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (Compound 9)¹H NMR(500MHz,MeOD)δ8.91(d,J＝2.5Hz,Ar-H,1H),8.38(dd,J＝8.5,2.5Hz,Ar-H, 1H),8.18(s,Ar-H,1H),8.03(d,J＝8.5Hz,Ar-H,1H),3.96–3.93(m,CH₂,2H),3.44(d,J＝7.0 Hz,CH₂,2H),3.36(td,J＝12.0,2.0Hz,CH₂,2H),2.12–2.01(m,CH,1H),1.70–1.66(m,CH₂, 2H),1.39–1.31(m,CH₂,2H)；ESI-MS：m/z＝397[M+1]⁺。

2.35- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide (Compound 10)¹H NMR(500MHz,DMSO-d₆)δ8.83(d,J＝2.5Hz,Ar-H,1H),8.46(dd,J＝8.5,2.5Hz, Ar-H,1H),8.26(s,Ar-H,1H),7.97(d,J＝8.5Hz,Ar-H,1H),7.94(s,NH₂-a,1H),7.48(s, NH₂-b,1H),6.65(d,J＝7.5Hz,NH,1H),4.04(m,CH,1H),2.58–2.56(m,CH,1H),1.86–1.82 (m,CH₂×2,4H),1.58–1.46(m,CH₂,2H),1.22–1.15(m,CH₂,2H)；ESI-MS：m/z＝396[M+1]⁺。

2.45- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide (Compound 11)¹H NMR(500MHz,DMSO-d₆)δ8.86(d,J＝2.5Hz,Ar-H,1H),8.42(dd,J＝8.5,2.5Hz, Ar-H,1H),8.30–8.22(m,Ar-H,1H),8.00–7.89(m,Ar-H,NH₂-a,2H),7.49(d,J＝3.0Hz, NH₂-b,1H),6.44–6.31(m,NH,1H),4.10(m,CH,1H),2.99–2.97(m,CH,1H),1.95–1.81(m, CH₂,2H),1.61–1.55(m,CH₂×2,4H),1.53–1.43(m,CH₂,2H)；ESI-MS：m/z＝396[M+1]⁺。

2.55- ((4- (((1r,4r) -4-methylcyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (Compound 12)¹H NMR(500MHz,DMSO-d₆)δ10.09(br,NH,1H),8.93–8.74(m,Ar-H,1H),8.44(dd,J ＝8.5,2.5Hz,Ar-H,1H),8.26(s,Ar-H,1H),7.95(d,J＝8.5Hz,Ar-H,1H),7.92(s,NH₂-a,1H), 7.48(s,NH₂-b,1H),6.64(d,J＝7.5Hz,NH,1H),4.05–4.02(m,CH,1H),1.87–1.85(m,CH₂, 2H),1.75–1.72(m,CH₂,2H),1.59–1.43(m,CH₂,2H),1.42–1.28(m,CH,1H),1.09–1.01(m, CH₂,2H),0.92(d,J＝6.5Hz,CH₃,3H)；ESI-MS：m/z＝395[M+1]⁺。

2.65- ((4- (((1r,4r) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (Compound 13)¹H NMR(500MHz,DMSO-d₆)δ10.11(br,NH,1H),8.82(d,J＝2.5Hz,Ar-H,1H),8.47 (dd,J＝8.5,2.5Hz,Ar-H,1H),8.26(s,Ar-H,1H),7.96(d,J＝8.5Hz,Ar-H,1H),7.92(d,J＝3.0 Hz,NH₂-a,1H),7.49(d,J＝3.0Hz,NH₂-b,1H),6.65(d,J＝7.5Hz,NH,1H),4.61(d,J＝5.0Hz, OH,1H),4.06–4.01(m,CH,1H),3.43–3.39(m,CH,1H),1.94–1.80(m,CH₂×2,4H),1.64– 1.45(m,CH₂,2H),1.33–1.26(m,CH₂,2H)；ESI-MS：m/z＝397[M+1]⁺。

2.75- ((4- (4-hydroxypiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide (Compound 14)¹H NMR(500MHz,DMSO-d₆)δ10.22(br,NH,1H),8.92(d,J＝2.5Hz,Ar-H,1H),8.47(s,Ar-H, 1H),8.31(dd,J＝8.5,2.5Hz,Ar-H,1H),7.98(d,J＝8.5Hz,Ar-H,NH₂-a,2H),7.63–7.38(m, NH₂-b,1H),4.83(d,J＝4.5Hz,OH,1H),3.88–3.83(m,CH₂,2H),3.80–3.74(m,CH,1H),3.32 –3.27(m,CH₂,2H),1.92–1.76(m,CH₂,2H),1.49–1.42(m,CH₂,2H)；ESI-MS：m/z ＝383[M+1]⁺。

2.85- ((4- (4-Aminopiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino)) picolinamide (Compound 15)¹H NMR(500MHz,DMSO-d₆)δ10.20(br,NH,1H),8.91(d,J＝2.5Hz,Ar-H,1H),8.46(s,Ar-H, 1H),8.31(dd,J＝8.5,2.5Hz,Ar-H,1H),8.02–7.93(m,Ar-H,NH₂-a,2H),7.46(d,J＝3.0Hz, NH₂-b,1H),4.02–3.98(m,CH₂,2H),3.17–3.11(m,CH₂,2H),2.87–2.83(m,CH,1H),1.84– 1.79(m,CH₂,2H),1.34–1.25(m,CH₂,2H)；ESI-MS：m/z＝382[M+1]⁺。

2.95- (4- (methylamino-5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (Compound 16)¹H NMR(500 MHz,DMSO-d₆)δ10.12(s,NH,1H),9.01(d,J＝2.5Hz,Ar-H,1H),8.36(dd,J＝8.5,2.5Hz, Ar-H,1H),8.25(s,Ar-H,1H),7.98(s,Ar-H,1H),7.96(s,NH₂-a,1H),7.46(d,J＝3.0Hz,NH₂-b, 1H),7.34(q,J＝4.5Hz,NH,1H),2.95(d,J＝4.5Hz,CH₃,3H)；ESI-MS：m/z＝313[M+1]⁺。

Preparation example 3 preparation of Compounds 17 to 19

Step 1, preparation of intermediate 1-21-1-22

Synthesis procedure referring to example 1, step 3, compounds 1-21 to 1-22 were prepared from the corresponding amines.

1.15-bromo-N-methylpyridinamide (intermediates 1-21)¹H NMR(500MHz,CDCl₃)δ8.58(d,J＝2.0 Hz,Ar-H,1H),8.08(d,J＝8.5Hz,Ar-H,1H),7.96(dd,J＝8.5,2.0Hz,Ar-H,1H),7.93–7.84(m, NH,1H),3.02(d,J＝5.0Hz,CH₃,3H).ESI-MS：m/z＝215[M+1]⁺。

1.25-bromo-N, N-dimethylpyridinamide (intermediate 1-22)¹H NMR(500MHz,CDCl₃)δ8.63(d, J＝2.0Hz,Ar-H,1H),7.92(dd,J＝8.5,2.0Hz,Ar-H,1H),7.57(dd,J＝8.5,2.0Hz,Ar-H,1H), 3.12(s,CH₃,3H),3.09(s,CH₃,3H).ESI-MS：m/z＝229[M+1]⁺。

Step 2. preparation of Compounds 17-19

Synthesis procedures refer to step 4 of preparation example 1, and 1-13/1-14 is used instead of 1-3, and 1-21 to 1-22/1-9 is used instead of 1-5 to 1-11, to prepare compounds 17 to 19.

2.1N- (5- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) pyridin-2-yl) cyclopropylcarboxamide (Compound 17)¹H NMR(500MHz,DMSO-d₆)δ10.08(br,NH,1H),8.79(s,Ar-H,1H), 8.52–8.49(m,Ar-H×2,2H),8.26(s,Ar-H,1H),7.95(d,J＝9.0Hz,Ar-H,1H),6.63(d,J＝8.0Hz, NH,1H),4.05(m,CH,1H),2.90–2.87(m,CH,1H),2.61–2.53(m,CH,1H),1.86–1.81(m, CH₂×2,4H),1.57–1.49(m,CH₂,2H),1.28–1.11(m,CH₂,2H),0.78–0.57(m,CH₂×2,4H)； ESI-MS：m/z＝436[M+1]⁺。

2.25- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylpyridinamide (Compound 18)¹H NMR(500MHz,DMSO-d₆)δ8.85(d,J＝2.5Hz,Ar-H,1H),8.56(q,J＝5.0Hz, NH,1H),8.40(dd,J＝8.5,2.5Hz,Ar-H,1H),8.28(s,Ar-H,1H),7.94(d,J＝8.5Hz,Ar-H,1H), 6.28(d,J＝7.5Hz,NH,1H),4.15–4.11(m,CH,1H),3.02–3.01(m,CH,1H),2.80(d,J＝5.0Hz, CH₃,3H),1.91–1.84(m,CH₂,2H),1.70–1.58(m,CH₂×2,4H),1.53–1.49(m,CH₂,2H)； ESI-MS：m/z＝410[M+1]⁺。

2.35- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylpyridine amide (Compound 19)¹H NMR(500MHz,DMSO-d₆)δ8.88(d,J＝2.5Hz,Ar-H,1H),8.28–8.26(m, Ar-H,1H),8.25(d,J＝2.5Hz,Ar-H,1H),7.54(d,J＝8.5Hz,Ar-H,1H),6.41–6.24(m,NH,1H), 4.11(s,CH,1H),3.02(s,CH₃,3H),2.99(s,CH₃,3H),1.90–1.83(m,CH₂,2H),1.62–1.55(m, CH₂×2,4H),1.53–1.47(m,CH₂,2H)；ESI-MS：m/z＝424[M+1]⁺。

Preparation example preparation of 45- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-cyclopropylpicolinamide (Compound 20)

Step 1 Synthesis of N- (5-bromopyridin-2-yl) cyclopropylcarboxamide (1-24)

Synthetic procedure referring to step 3 of preparation example 1, intermediates 1-24 were prepared using 1-23 instead of amine and cyclopropanecarboxylic acid instead of acid. Yield: 60 percent;¹H NMR(500MHz,CDCl₃)δ9.32(br,NH,1H),8.25(d,J＝2.5Hz,Ar-H,1H),8.19 (d,J＝9.0Hz,Ar-H,1H),7.79(dd,J＝9.0,2.5Hz,Ar-H,1H),1.66–1.60(m,CH,1H),1.10– 1.07(m,CH₂,2H),0.91–0.88(m,CH₂,2H).ESI-MS：m/z＝241[M+1]⁺。

step 2 Synthesis of Compound 5- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-cyclopropylpicolinamide (Compound 20)

Synthesis procedure with reference to step 4 of preparation example 1, 1-13 was used in place of 1-3, and 1-24 was used in place of 1-5 to 1-11, to prepare Compound 20. Yield: 30 percent;¹H NMR(500MHz,DMSO-d₆)δ10.69(br,NH,1H),9.70(br,NH,1H),8.73 –8.52(m,Ar-H,1H),8.19(s,Ar-H,1H),8.07(dd,J＝9.0,2.5Hz,Ar-H,1H),7.99(d,J＝9.0Hz, Ar-H,1H),6.44(d,J＝8.0Hz,NH,1H),4.03–4.00(m,CH,1H),2.67–2.57(m,CH,1H),2.01– 1.96(m,CH,1H),1.91–1.75(m,CH₂×2,4H),1.59–1.45(m,CH₂,2H),1.18–1.04(m,CH₂,2H), 0.82–0.75(m,CH₂×2,4H)；ESI-MS：m/z＝436[M+1]⁺。

preparation examples 5 preparation of Compounds 21 to 26

Step 1 Synthesis of Compound (1s,4s) -4- ((2-amino-5-trifluoromethylpyrimidin-4-yl) amino) cyclohexanol (1-25)

Synthetic procedure referring to preparation example 1, step 2, compounds 1-25 were prepared using the corresponding amine instead of 1-Boc-4-aminomethylpiperidine. The yield is 75 percent; ESI-MS: m/z 277[ M + 1]]⁺。

Step 2, synthesizing a compound 21-26

Synthesis procedures referring to step 4 of preparation example 1, compounds 21 to 26 were prepared by substituting 1-13/1-14/1-16/1-25/1-17/1-18 for 1-3 and 1-26 for 1-5 to 1-11. Yield: 40-60 percent;

2.14- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 21)¹H NMR(500MHz,DMSO-d₆)δ9.91(s,NH,1H),8.22(s,Ar-H,1H),7.87–7.81(m, Ar-H×4,NH₂-a,5H),7.16(s,NH₂-b,1H),6.57(d,J＝6.5Hz,NH,1H),4.07–4.05(m,CH,1H), 2.73–2.69(m,CH,1H),1.94–1.86(m,CH₂×2,4H),1.60–1.45(m,CH₂,2H),1.45–1.26(m, CH₂,2H)；ESI-MS：m/z＝395[M+1]⁺。

2.24- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 22)¹H NMR(500MHz,DMSO-d₆)δ9.92(br,NH,1H),8.25(s,Ar-H,1H),7.85(s,NH₂-a,1H), 7.82(s,Ar-H×4,4H),7.21(s,,NH₂-b,1H),6.28(d,J＝7.0Hz,NH,1H),4.14–4.12(m,CH,1H), 3.06–3.02(m,CH,1H),1.92–1.85(m,CH₂,2H),1.66–1.58(m,CH₂×2,4H),1.57–1.52(m, CH₂,2H)；ESI-MS：m/z＝395[M+1]⁺。

2.34- ((4- (((1r,4r) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 23)¹H NMR(500MHz,DMSO-d₆)δ9.90(br,NH,1H),8.22(s,Ar-H,1H),7.86(s,NH₂-a,1H), 7.82(s,Ar-H,4H),7.21(s,NH₂-b,1H),6.58(d,J＝7.5Hz,NH,1H),4.06(m,CH,1H),3.44– 3.38(m,CH,1H),1.96–1.79(m,CH₂×2,4H),1.57–1.49(m,CH₂,2H),1.36–1.25(m,CH₂,2H)； ESI-MS：m/z＝396[M+1]⁺。

2.44- ((4- (((1s,4s) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 24)¹H NMR(500MHz,DMSO-d₆)δ9.90(br,NH,1H),8.23(s,Ar-H,1H),7.82(m, Ar-H+NH₂-a,5H),7.20(s,NH₂-b,1H),6.50(d,J＝7.5Hz,NH,1H),4.43(d,J＝3.0Hz,OH,1H), 4.15–4.04(m,CH,1H),3.87–3.85(m,CH,1H),1.91–1.83(m,CH₂,2H),1.76–1.68(m,CH₂, 2H),1.68–1.60(m,CH₂,2H),1.59–1.53(m,CH₂,2H)；ESI-MS：m/z＝396[M+1]⁺。

2.54- ((4- (4-hydroxypiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 25)¹H NMR(500MHz,DMSO-d₆)δ9.98(br,NH,1H),8.43(s,Ar-H,1H),7.87–7.74(m,Ar-H×4, NH₂-a,5H),7.26–7.12(m,NH₂-b,1H),4.81(d,J＝4.5Hz,OH,1H),3.89–3.85(m,CH₂,2H), 3.80–3.74(m,CH,1H),3.32–3.26(m,CH₂,2H),1.88–1.83(m,CH₂,2H),1.50–1.43(m,CH₂, 2H)；ESI-MS：m/z＝382[M+1]⁺。

2.64- ((4- (4-Aminopiperidin-1-yl) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 26)¹H NMR(500MHz,DMSO-d₆)δ9.98(br,NH,1H),8.43(s,Ar-H,1H),7.88–7.72(m,Ar-H×4, NH₂-a,5H),7.19(s,NH₂-b,1H),4.04–3.99(m,CH₂,2H),3.16–3.11(m,CH₂,2H),2.89–2.83 (m,CH,1H),1.84–1.79(m,CH₂,2H),1.34–1.27(m,CH₂,2H)；ESI-MS：m/z＝381[M+1]⁺。

Preparation of Compounds 27 to 28 of preparation example 6

Step 1, synthesizing compounds 27-28

Synthesis procedure with reference to step 4 of preparation example 1, compounds 27 to 28 were prepared by substituting 1-14 for 1-3 and 1-27/1-28 for 1-5 to 1-11. Yield: 50-60%;

1.14- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylbenzamide (Compound 27)¹H NMR(500MHz,DMSO-d₆)δ9.68(br,NH,1H),8.22(s,Ar-H,1H),7.63(s, Ar-H,1H),7.56(d,J＝8.5Hz,Ar-H,NH₂-a,2H),7.34(d,J＝8.5Hz,Ar-H,1H),7.13(s,NH₂-b, 1H),6.12(d,J＝7.5Hz,NH,1H),4.18–4.13(m,CH,1H),3.01–2.94(m,CH,1H),2.38(s,CH₃, 3H),1.94–1.81(m,CH₂,2H),1.64–1.56(m,CH₂×2,4H),1.52–1.46(m,CH₂,2H)；.ESI-MS： m/z＝409[M+1]⁺。

1.24- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methoxybenzamide (Compound 28)¹H NMR(500MHz,DMSO-d₆)δ9.87(s,NH,1H),8.26(s,Ar-H,1H),7.80(d,J＝ 8.5Hz,Ar-H,1H),7.59–7.57(dd,J＝8.5,2.0Hz,1H),7.53(s,NH₂-a,1H),7.47(s,Ar-H,1H), 7.41(s,NH₂-b,1H),6.26(d,J＝7.5Hz,NH,1H),4.22–4.15(m,CH,1H),3.89(s,CH₃,3H),2.98 –2.95(m,CH,1H),1.95–1.79(m,CH₂,2H),1.62–1.53(m,CH₂×2,4H),1.50–1.45(m,CH₂, 2H)；ESI-MS：m/z＝425[M+1]⁺。

Preparation of Compounds 29 to 32 of preparation example 7

Step 1, synthesizing an intermediate 1-30-1-32

Synthesis procedure referring to step 3 of preparation example 1, intermediates 1-30 to 1-32 were prepared using the corresponding amine, substituting 1-29 for the acid. Yield: 70-90%.

1.14 Synthesis of bromo-N-methylbenzamide (intermediate 1-30)¹H NMR(500MHz,CDCl₃)δ7.65– 7.60(m,Ar-H,2H),7.57–7.53(m,Ar-H,2H),6.35–6.17(m,NH,1H),2.99(d,J＝5.0Hz,CH₃, 3H).ESI-MS：m/z＝214[M+1]⁺。

Synthesis of 24-bromo-N, N-dimethylbenzamide (intermediate 1-31)¹H NMR(500MHz,CDCl₃)δ7.55 –7.51(m,Ar-H,2H),7.31–7.27(m,Ar-H,2H),3.08(s,CH₃,3H),2.97(s,CH₃,3H).ESI-MS： m/z＝228[M+1]⁺。

1.31- (4-bromobenzoyl) piperidine (intermediate 1-32) Synthesis¹H NMR(500MHz,DMSO-d₆)δ7.65– 7.61(m,Ar-H,2H),7.34–7.30(m,Ar-H,2H),3.56(s,CH₂,2H),3.23(s,CH₂,2H),1.62–1.58(m, CH₂,2H),1.53-1.44(m,CH₂×2,4H).ESI-MS：m/z＝268[M+1]⁺。

Step 2, synthesizing compounds 29-32

The synthetic procedure refers to step 4 of preparation example 1, 1-14/1-25 is used instead of 1-3, and 1-30-1-32 is used instead of 1-5-1-11, to prepare compounds 29-32. Yield: 40-60%;

2.14- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylbenzamide (Compound 29)¹H NMR(500MHz,DMSO-d₆)δ9.85(br,NH,1H),8.25–8.24(m,Ar-H,NH,2H), 7.82(d,J＝8.5Hz,Ar-H,2H),7.76(d,J＝8.5Hz,Ar-H,2H),6.21(d,J＝7.5Hz,NH,1H),4.16– 4.12(m,CH,1H),3.04–2.98(m,CH,1H),2.77(d,J＝5.0Hz,CH₃,3H),1.88(q,J＝10.0Hz, CH₂,2H),1.70–1.56(m,CH₂×2,4H),1.55–1.50(m,CH₂,2H)；ESI-MS：m/z＝409[M+1]⁺。

2.24- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylbenzamide (Compound 30)¹H NMR(500MHz,DMSO-d₆)δ9.86(br,NH,1H),8.23(s,Ar-H,1H),7.80(d, J＝8.5Hz,Ar-H,2H),7.34(d,J＝8.5Hz,Ar-H,2H),6.24(d,J＝7.5Hz,NH,1H),4.13(m,CH, 1H),3.00–2.99(m,CH,1H),2.95(s,CH₃×2,6H),1.95–1.78(m,CH₂,2H),1.67–1.55(m, CH₂×2,4H),1.55–1.46(m,CH₂,2H)；ESI-MS：m/z＝423[M+1]⁺。

2.34- ((4- (((1s,4s) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylbenzamide (Compound 31)¹H NMR(500MHz,DMSO-d₆)δ9.85(br,NH,1H),8.22(s,Ar-H,1H),7.80(d, J＝8.0Hz,Ar-H,2H),7.34(d,J＝8.0Hz,Ar-H,2H),6.48(d,J＝7.5Hz,NH,1H),4.43(s,OH, 1H),4.08–4.07(m,CH,1H),3.92–3.73(m,CH,1H),2.95(s,CH₃×2,6H),1.94–1.78(m,CH₂, 2H),1.74–1.65(m,CH₂,2H),1.65–1.58(m,CH₂,2H),1.56–1.49(m,CH₂,2H)；ESI-MS：m/z＝ 424[M+1]⁺。

2.44- ((4- (((1s,4s) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) (piperidin-1-yl) methanone (Compound 32)¹H NMR(500MHz,DMSO-d₆)δ9.83(s,NH,1H),8.21(s,Ar-H,1H),7.85– 7.75(m,Ar-H,2H),7.35–7.25(m,Ar-H,2H),6.46(d,J＝7.0Hz,NH,1H),4.42(d,J＝3.0Hz, OH,1H),4.17–3.99(m,CH,1H),3.84–3.82(m,CH,1H),3.46-3.41(m,CH₂×2,1H),1.89–1.81 (m,CH₂,2H),1.70–1.67(m,CH₂,2H),1.64–1.58(m,CH₂×2,4H),1.57–1.44(m,CH₂×3,6H)； ESI-MS：m/z＝464[M+1]⁺。

Preparation example 8 preparation of Compounds 33 to 36

Step 1. Synthesis of intermediates 1 to 34

Synthetic procedure referring to step 3 of preparation example 1, intermediates 1-34 were prepared in yields using the corresponding acids, substituting 1-33 for the amine: 70-80%.

1.1 Synthesis of N- (4-bromobenzene) cyclopropylcarboxamide (intermediate 1-34)¹H NMR(500MHz,CDCl₃)δ7.93 (br,NH,1H),7.42(d,J＝9.0Hz,Ar-H,2H),7.38(d,J＝9.0Hz,Ar-H,2H),1.57–1.49(m,CH, 1H),1.09–1.03(m,CH₂,2H),0.85–0.79(m,CH₂,2H).ESI-MS：m/z＝241[M+1]⁺。

Step 2, synthesizing a compound 33-36

Synthesis procedures referring to step 4 of example 1, compounds 33 to 36 were prepared by replacing 1-3 with 1-13/1-14/1-25 and 1-5 to 1-11 with 1-34 to 1-35. Yield: 30-40%.

2.1N- (4- ((4- (((1r,4r) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) cyclopropylcarboxamide (Compound 33)¹H NMR(500MHz,DMSO-d₆)δ10.12(br,NH,1H),9.57(br,NH,1H),8.15 (s,Ar-H,1H),7.64(d,J＝8.5Hz,Ar-H,2H),7.49(d,J＝8.5Hz,Ar-H,2H),6.39(d,J＝7.5Hz, NH,1H),4.21–3.86(m,CH,1H),2.65–2.59(m,CH,1H),1.88–1.83(m,CH₂×2,4H),1.76– 1.72(m,CH,1H),1.51(m,CH₂,2H),1.19–1.12(m,CH₂,2H),0.88–0.67(m,CH₂×2,4H)； ESI-MS：m/z＝435[M+1]⁺。

2.2N- (4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) acetamide (Compound 34)¹H NMR(500MHz,DMSO-d₆)δ9.84(s,NH,1H),9.57(br,NH,1H),8.17(s,Ar-H, 1H),7.71–7.55(m,Ar-H,2H),7.53–7.41(m,Ar-H,2H),6.13(br,NH,1H),4.09(m,CH,1H), 3.00–2.95(m,CH,1H),2.01(s,CH₃,3H),1.89–1.81(m,CH₂,2H),1.64–1.57(m,CH₂,2H), 1.56–1.49(m,CH₂×2,4H)；ESI-MS：m/z＝409[M+1]⁺。

2.3N- (4- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) cyclopropylcarboxamide (Compound 35)¹H NMR(500MHz,DMSO-d₆)δ10.09(s,NH,1H),9.57(br,NH,1H),8.17(s, Ar-H,1H),7.63(d,J＝9.0Hz,Ar-H,2H),7.48(d,J＝9.0Hz,Ar-H,2H),6.12(br,NH,1H),4.26 –3.92(m,CH,1H),3.01–2.95(m,CH,1H),1.89–1.81(m,CH₂,2H),1.74(td,J＝8.0,4.0Hz, CH,1H),1.64–1.57(m,CH₂,2H),1.56–1.49(m,CH₂×2,4H),0.80–0.73(m,CH₂×2,4H)； ESI-MS：m/z＝435[M+1]⁺。

2.4N- (4- ((4- (((1s,4s) -4-Hydroxycyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -phenyl) acetamide (Compound 36)¹H NMR(500MHz,DMSO-d₆)δ9.83(s,NH,1H),9.73–9.43(m,NH,1H),8.16(s, Ar-H,1H),7.67–7.60(m,Ar-H,2H),7.50–7.42(m,Ar-H,2H),6.33(s,NH,1H),4.43(d,J＝3.0 Hz,OH,1H),4.05(m,CH,1H),3.82(m,CH,1H),2.00(s,CH₃,3H),1.88–1.80(m,CH₂,2H), 1.70–1.68(m,CH₂,2H),1.66–1.57(m,CH₂,2H),1.53–1.47(m,CH₂,2H)；ESI-MS：m/z＝ 410[M+1]⁺。

Preparation of Compounds 37 to 42 of preparation example 9

Step 1, synthesizing an intermediate 1-36-1-39

Synthesis procedure referring to step 2 of preparation example 1, compounds 1-36 to 1-39 were prepared by substituting 1-Boc-4-aminomethylpiperidine with the corresponding amine. The yield is 40-80%.

1.1 1-Boc-N⁴- ((1S,2S,4R) -4-Aminofluorocyclohexyl-5-trifluoromethylpyrimidine-2, 4-diamine (intermediate 1-36) LCMS: M/z ═ 394[ M + 1] M]⁺。

1.2 1-Boc-N⁴-8-aminobicyclo [3.2.1]Octane-3-yl-5-trifluoromethylpyrimidine-2, 4-diamine (intermediate 1-37) LCMS: m/z is 402[ M + 1]]⁺。

1.3 1-Boc-N⁴-8-Aminoazaspiro [4.5]]Decane-5-trifluoromethylpyrimidine-2, 4-diamine (intermediate 1-38) ESI-MS: m/z 416[ M + 1]]⁺。

1.4N⁴- (1-methyl-1H-pyrazol-4-yl) -5-trifluoromethylpyrimidine-2, 4-bisAmine (intermediates 1-39)¹H NMR(500 MHz,DMSO-d₆)δ8.55(s,Ar-H,1H),8.24(s,Ar-H,1H),8.14–8.06(m,NH,1H),7.73(s,Ar-H, 1H),7.00(s,NH×2,2H),3.79(s,CH₃,3H).ESI-MS：m/z＝259[M+1]⁺。

Step 2, synthesizing a compound 37-42

Synthesis procedures referring to step 4 of example 1, compounds 37 to 42 were prepared by substituting 1-36 to 1-39 for 1-3 and 1-26 or 1-20 for 1-5 to 1-11. Yield: 30-50%.

2.14- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 37), ESI-MS: m/z 413[ M + 1[ ]]⁺。

2.24- ((4- ((3-Aminobicyclo [ 3.2.1)]Oct-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 38), ESI-MS: 421[ M + 1] M/z]⁺。

2.34- ((4- ((2-azaspiro [ 4.5)]Decan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 39), ESI-MS: 435[ M + 1] M/z]⁺。

2.44- ((4- ((1-methyl-1H-pyrazol-4-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 40), ESI-MS: 378[ M + 1] M/z]⁺。

2.55- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (compound 41), ESI-MS: m/z 414[ M + 1]]⁺。

2.65- ((4- ((3-Aminobicyclo [ 3.2.1)]Octane-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) picolinamide (compound 42), ESI-MS: 422[ M + 1] M/z]⁺。

Preparation example 106- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) isoquinolineacetic acid methyl ester-1 (2H) -one (Compound 43)

Step 1 Synthesis of Compound 6- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) isoquinolineacetic acid methyl ester-1 (2H) -one (Compound 43)

Synthetic procedure referring to preparation example 1, step 4, compound 43 was prepared by substituting 1-14 for 1-3 and 2-1 for 1-5 to 1-11. Yield: 30 percent;¹H NMR(500MHz,DMSO-d₆)δ10.06(s,NH,1H),8.28(s,Ar-H,1H),8.09(d, J＝2.0Hz,Ar-H,1H),8.05(d,J＝8.5Hz,Ar-H,1H),7.73(dd,J＝8.5,2.0Hz,Ar-H,1H),7.12(d, J＝7.0Hz,CH,1H),6.35(d,J＝7.0Hz,CH,1H),6.29(d,J＝7.5Hz,NH,1H),4.20–4.16(m, CH,1H),3.06–2.95(m,CH,1H),2.02–1.80(m,CH₂,2H),1.68–1.56(m,CH₂×2,4H),1.54– 1.49(m,CH₂,2H)；ESI-MS：m/z＝419[M+1]⁺。

preparation example 116 preparation of methyl- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolineacetate-1 (2H) -one (Compound 44)

Step 1. Synthesis of intermediate 6-bromo-2-methylisoquinolin-1 (2H) -one 2-2

The compound 2-16-bromo-2H-isoquinolin-1-one (224mg,1mmol) was dissolved in 2mL DMF and CS was added sequentially₂CO₃(650mg,2mmol), potassium iodide (170mg,1.2mmol), at 50 ℃ for 3 h. After cooling to room temperature, 10mL of water was added, EA extraction (10 mL. times.3) was performed, the organic layers were combined, washed with water (30 mL. times.1) and saturated brine (30 mL. times.1), dried over anhydrous sodium sulfate, the solvent was removed by distillation under the reduced pressure, and the mixture was purified by silica gel column chromatography using PE: EA (2:1) as an eluent, to obtain a white solid. Yield: 90 percent;¹H NMR(500MHz,DMSO-d₆)δ8.10(d,J＝8.5Hz,Ar-H,1H), 7.94(d,J＝2.0Hz,Ar-H,1H),7.62(dd,J＝8.5,2.0Hz,Ar-H,1H),7.53(d,J＝7.5Hz,CH,1H), 6.58(d,J＝7.5Hz,CH,1H),3.49(s,CH₃,3H).ESI-MS：m/z＝239[M+1]⁺。

step 2.6 Synthesis of methyl- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolineacetate-1 (2H) -one (Compound 44)

Synthesis procedure with reference to step 4 of example 1, compound 44 was prepared by substituting 1-14 for 1-3 and 2-2 for 1-5 to 1-11. Yield: 50 percent;¹H NMR(500MHz,DMSO-d₆)δ10.06(s,NH,1H),8.27(s,Ar-H,1H),8.09(dd,J＝ 5.5,3.5Hz,Ar-H,2H),7.74(dd,J＝9.0,2.0Hz,Ar-H,1H),7.41(d,J＝7.5Hz,CH,1H),6.41(d, J＝7.5Hz,CH,1H),6.29(d,J＝7.5Hz,NH,1H),4.22–4.15(m,CH,1H),3.46(s,CH₃,3H), 3.00–2.97(m,CH,1H),1.93–1.86(m,CH₂,2H),1.65–1.55(m,CH₂×2,4H),1.52–1.47(m, CH₂,2H)；ESI-MS：m/z＝433[M+1]⁺。

preparation example preparation of methyl- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2- (2-hydroxyethyl) isoquinolineacetate-1 (2H) -one 126 (Compound 45)

Step 1. Synthesis of intermediate 6-bromo-2- (2-hydroxyethyl) isoquinolin-1 (2H) -one 2-3

Synthetic procedure referring to example 11, step 1, compound 2-3 was prepared by substituting 1-14 for 1-3 and bromopropanol for iodomethane. Yield: 90 percent;¹H NMR(500MHz,DMSO-d₆)δ8.11(d,J＝8.5Hz,Ar-H,1H),7.93(d,J＝2.0Hz, Ar-H,1H),7.62(dd,J＝8.5,2.0Hz,Ar-H,1H),7.46(d,J＝7.5Hz,CH,1H),6.56(d,J＝7.5Hz, CH,1H),4.89(t,J＝5.5Hz,OH,1H),4.00(t,J＝5.5Hz,CH₂,2H),3.66(q,J＝5.5Hz,CH₂,2H). ESI-MS：m/z＝268[M+1]⁺。

step 2.6 Synthesis of methyl- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2- (2-hydroxyethyl) isoquinolineacetate-1 (2H) -one (Compound 45)

Synthetic procedure with reference to example 1, step 4, compound 45 was prepared by substituting 1-14 for 1-3 and 2-3 for 1-5 to 1-11. Yield: 50 percent;¹H NMR(500MHz,DMSO-d₆)δ10.06(s,NH,1H),8.27(s,Ar-H,1H),8.16–8.03(m, Ar-H,2H),7.73(dd,J＝9.0,2.0Hz,Ar-H,1H),7.36(d,J＝7.5Hz,CH,1H),6.39(d,J＝7.5Hz, CH,1H),6.28(d,J＝7.5Hz,NH,1H),4.91(s,OH,1H),4.21–4.16(m,CH,1H),3.97(t,J＝5.5 Hz,CH₂,2H),3.65(t,J＝5.5Hz,CH₂,2H),2.99–2.97(m,CH,1H),1.93–1.86(m,CH₂,2H), 1.65–1.56(m,CH₂×2,4H),1.53–1.47(m,CH₂,2H)；ESI-MS：m/z＝477[M+1]⁺。

preparation example 136- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-cyclopropanemethylisoquinolineacetic acid methyl ester-1 (2H) -one (Compound 46)

Step 1.6-bromo-2- (2-cyclopropylmethyl) isoquinolin-1 (2H) -one 2-4 Synthesis

Synthetic procedure referring to example 11, step 1, using cyclopropylmethylThe methyl iodide is replaced by the chlorine radical to prepare an intermediate 2-4. Yield: 90 percent;¹H NMR(500MHz,DMSO-d₆)δ8.12(d,J＝8.5Hz,Ar-H,1H),7.94(d,J＝2.0Hz,Ar-H,1H), 7.63(dd,J＝8.5,2.0Hz,Ar-H,1H),7.58(d,J＝7.5Hz,CH,1H),6.60(d,J＝7.5Hz,CH,1H), 3.81(d,J＝7.0Hz,CH₂,2H),1.29–1.16(m,CH,1H),0.50–0.45(m,CH₂,2H),0.41–0.38(m, CH₂,2H).ESI-MS：m/z＝278[M+1]⁺。

step 2.6 Synthesis of methyl- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-cyclopropanemethylisoquinolineacetate-1 (2H) -one (Compound 46)

Synthesis procedure with reference to step 4 of example 1, compound 46 was prepared by substituting 1-14 for 1-3 and 2-4 for 1-5 to 1-11. Yield: 50 percent;¹H NMR(500MHz,DMSO-d₆)δ10.08(br,NH,1H),8.28(s,Ar-H,1H),8.15–8.07 (m,Ar-H,2H),7.75(dd,J＝9.0,2.0Hz,Ar-H,1H),7.47(d,J＝7.5Hz,CH,1H),6.42(d,J＝7.5 Hz,CH,1H),6.30(d,J＝7.5Hz,NH,1H),4.23–4.16(m,CH,1H),3.78(d,J＝7.0Hz,CH₂,2H), 3.00–2.97(m,CH,1H),1.94–1.87(m,CH₂,2H),1.66–1.56(m,CH₂×2,4H),1.54–1.48(m, CH₂,2H),1.25–1.18(m,CH,1H),0.49–0.45(m,CH₂,2H),0.41–0.36(m,CH₂,2H)；ESI-MS： m/z＝473[M+1]⁺。

preparation example 146- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-difluoromethylisoquinolin-1 (2H) -one (Compound 47)

Synthesis procedure with reference to step 4 of example 1, compound 47 was prepared by substituting 1-14 for 1-3 and 2-5 for 1-5 to 1-11. Yield: 50 percent; ESI-MS: m/z 469[ M +1 ═ M]⁺。

Preparation example 156- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -1-oxoisoquinoline-2 (1H) -benzonitrile (Compound 48)

Synthesis procedure with reference to step 4 of example 1, compound 47 was prepared by substituting 1-14 for 1-3 and 2-6 for 1-5 to 1-11. Yield: 50 percent; ESI-MS: 444[ M + 1] M/z]⁺。

Preparation example 166- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-difluoromethylisoquinolin-1 (2H) -one (Compound 49)

Synthesis procedure with reference to step 4 of example 1, compound 49 was prepared by substituting 1-14 for 1-3 and 2-7 for 1-5 to 1-11. Yield: 50 percent; ESI-MS: m/z 459[ M + 1%]⁺。

Preparation of Compounds 50 to 52 of preparation example 17

Synthesis procedures referring to step 4 of example 1, compounds 50 to 52 were prepared by substituting 1-36 to 1-38 for 1-3 and 2-2 for 1-5 to 1-11. Yield: 40-50%.

6- ((4- (((1R,3S,4S) -4-amino-3-fluorocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one (Compound 50) ESI-MS: 451[ M + 1] M/z]⁺。

6- ((4- ((3-Aminobicyclo [ 3.2.1)]Octane-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolin-1 (2H) -one (Compound 51) ESI-MS: m/z 459[ M + 1]]⁺。

6- ((4- ((2-azaspiro [ 4.5)]Decan-8-yl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylIsoquinolin-1 (2H) -one (compound 52) ESI-MS: m/z 473[ M +1 ═ M]⁺。

Preparation example 185- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) isoindolin-1-one (Compound 53)

Synthesis procedure with reference to step 4 of example 1, compound 53 was prepared by substituting 1-14 for 1-3 and 2-8 for 1-5 to 1-11. Yield: 40-50%.¹H NMR(500MHz,DMSO-d₆)δ10.01(br,NH,1H),8.33(s,Ar-H,1H),8.25(s, NH,1H),8.09(d,J＝2.0Hz,Ar-H,1H),7.74(dd,J＝8.5,2.0Hz,Ar-H,1H),7.55(d,J＝8.5Hz, Ar-H,1H),6.30(d,J＝7.5Hz,NH,1H),4.33(s,CH₂,2H),4.16–4.10(m,CH₂,1H),3.03–2.96 (m,CH,1H),1.97–1.81(m,CH₂,2H),1.66–1.46(m,CH₂×3,6H)；ESI-MS：m/z＝407[M+1]⁺。

Preparation example 195- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoindolin-1-one (Compound 54)

Step 1. Synthesis of intermediate 5-bromo-2-methyl-1, 3-dihydroisoindol-1-one 2-10

Synthetic procedures intermediates 2-10 were prepared by substituting 2-9 for 2-1, in reference to example 11, step 1. Yield: 90 percent;¹H NMR (500MHz,CDCl₃)δ7.70–7.67(m,Ar-H,1H),7.61–7.56(m,Ar-H,2H),4.35(s,CH₂,2H),3.18 (s,CH₃,3H).ESI-MS：m/z＝226[M+1]⁺。

step 2.5- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoindolin-1-one (Compound 54) Synthesis

Synthesis procedure with reference to step 4 of example 1, compound 54 was prepared by substituting 1-14 for 1-3 and 2-10 for 1-5 to 1-11. Yield: 50 percent;¹H NMR(500MHz,DMSO-d₆)δ10.01(br,NH,1H),8.25(s,Ar-H,1H),8.07(s,Ar-H, 1H),7.74(dd,J＝8.5,2.0Hz,Ar-H,1H),7.54(d,J＝8.5Hz,Ar-H,1H),6.27(d,J＝7.5Hz,NH, 1H),4.41(s,CH₂,2H),4.16–4.10(m,CH,1H),3.04(s,CH₃,3H),3.02–3.01(m,CH,1H),1.94– 1.81(m,CH₂,2H),1.66–1.56(m,CH₂×2,4H),1.55–1.47(m,CH₂,2H)；ESI-MS：m/z＝ 421[M+1]⁺。

preparation example 20N- (5- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino)) pyridin-2-yl) methanesulfonamide (Compound 55)

Synthesis procedure with reference to step 4 of example 1, compound 55 was prepared by substituting 1-14 for 1-3 and 2-11 for 1-5 to 1-11. Yield: 50 percent. ESI-MS: m/z 446[ M + 1]]⁺。

Preparation example 215- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-methylpyridine-2-sulphonamide (compound 56)

Synthesis procedure with reference to step 4 of example 1, compound 56 was prepared by substituting 1-14 for 1-3 and 2-12 for 1-5 to 1-11. Yield: 50 percent. ESI-MS: m/z 446[ M + 1]]⁺。

Preparation example 224- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-toluenesulfonamide (Compound 57)

Synthesis procedure with reference to step 4 of example 1, compound 57 was prepared by substituting 1-14 for 1-3 and 2-13 for 1-5 to 1-11. Yield: 50 percent. ESI-MS: m/z 445[ M + 1]]⁺。

Preparation example 234- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N-toluenesulfonamide (Compound 58)

Synthesis procedure with reference to step 4 of example 1, compound 57 was prepared by substituting 1-14 for 1-3 and 2-14 for 1-5 to 1-11. Yield: 50 percent. ESI-MS: m/z 445[ M + 1]]⁺。

Preparation example 244- ((4- (((1s,4s) -4-Aminocyclohexyl ether) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (compound 54)

Step 1 Synthesis of intermediate 1-Boc- ((1s,4s) -4- ((2-amino-5-trifluoromethylpyrimidin-4-yl) cyclopropyl ether 3-1

Compound 1-2(198mg,1.0mmol) was dissolved in anhydrous tetrahydrofuran (5mL), sodium hydride (60mg,1.5mmol) was added in portions with stirring in an ice bath, followed by cis-N-BOC-4-aminocyclohexanol (258mg,1.2 mmol/1 mL THF), reacted for 15min in an ice bath, and then moved to room temperature for 10 h. After quenching with saturated ammonium chloride solution, 10mL of water was added, EA extraction (10 mL. times.3) was carried out, the organic layer was separated, and the mixture was washed with saturated brine (20 mL. times.1) and anhydrous sulfuric acidDrying sodium, distilling under reduced pressure to remove solvent, and purifying with silica gel column chromatography with PE: EA (3:1) as eluent to obtain white solid 3-1. Yield: 30 percent; ESI-MS: m/z 377[ M +1 ═ M]⁺。

Step 2.Synthesis of 4- ((4- (((1s,4s) -4-aminocyclohexyl ether) -5-trifluoromethylpyrimidin-2-yl) amino) benzamide (Compound 59)

Synthesis procedure with reference to step 4 of example 1, compound 59 was prepared by substituting 3-1 for 1-3 and 3-2 for 1-5 to 1-11. Yield: 30 percent; ESI-MS: 396[ M + 1] M/z]⁺。

Preparation example 254- ((4- (((1s,4s) -4-aminocyclohexane) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -N, N-dimethylbenzamide salt-forming Compound 60-73

Citric acid, fumaric acid, malic acid, L-malic acid, D-malic acid, methanesulfonic acid, L-tartaric acid, D-tartaric acid, succinic acid, maleic acid, formic acid, acetic acid, hydrochloric acid, phosphoric acid

Dissolving 1.2 equivalents of organic acid or inorganic acid in an ethanol solution, slowly dripping the solution into the ethanol solution of the compound 30, adding a proper amount of ether solution, carrying out suction filtration to obtain a salt-forming compound, washing with ether, and drying to obtain the corresponding salt-forming compound 60-73.

Preparation example 266- ((4- (((1s,4s) -4-aminocyclohexyl) amino) -5-trifluoromethylpyrimidin-2-yl) amino) -2-methylisoquinolineacetic acid methyl ester-1 (2H) -one salt-forming compound 74-87

Dissolving 1.2 equivalents of organic acid or inorganic acid in an ethanol solution, slowly and dropwise adding the solution into the ethanol solution of the compound 44, adding a proper amount of an ether solution, carrying out suction filtration to obtain a salt-forming compound, washing with ether, and drying to obtain the corresponding salt-forming compound 74-87.

The following examples describe the biological activity of the compounds of the present invention, but the present invention is not limited to these examples.

Example 27 testing of inhibition of FLT3, FLT3-D835Y, c-KIT kinase

The method adopts specific biotin-labeled polypeptide TK Substrate as a Substrate, and the reaction is started by FLT3, FLT3-D835Y and c-KIT in the presence of ATP, so that phosphorylation modification of the polypeptide TK Substrate can be catalyzed. The Eu-labeled TK Substrate-specific phosphorylated antibody can be bound to the Substrate by antigen-antibody reaction, while the streptavidin-labeled receptor is bound by the specific interaction of streptavidin with biotin. Thereby enabling the Eu-labeled donor to interact with the streptavidin-labeled receptor. In fluorescence resonance energy transfer, when two fluorophores are close due to biomolecular interaction, part of the energy captured by the cryptate will be released upon excitation, with an emission wavelength of 620 nm; another part of the energy is transferred to the acceptor (acceptor), the emission wavelength is 665 nm. The emission at 665nm is generated only by FRET by the donor (donor). Therefore, when biomolecules interact, there are two excitation lights 620nm and 665 nm; when there is no interaction, there is only 620nm excitation light. FLT3 phosphorylation activity can be reflected by measuring the ratio of the fluorescence signals at two emission wavelengths of 665nm and 620 nm. Meanwhile, a blank control is arranged to judge the strength of the enzyme activity. Giltertinib was used as a positive inhibitor in the experiment.

The samples were dissolved in DMSO and stored at low temperature, and the concentration of DMSO in the final system was controlled within a range that did not affect the detection activity. FLT3, FLT3-D835Y and substrate were treated with HTRF Kinase buffer (1.25X Kinase buffer,6.25mM MgCl. RTM₂,1.25mM MnCl₂1.25mM DTT). Will be provided withmu.L of enzyme, 4. mu.L of substrate and 2. mu.L of compound to be tested at different concentrations are added to a 384 reaction plate (ProxiPlate TM-384Plus, Perkinelmer) respectively, and the specific reaction system is 2% DMSO, 0.5 ng/. mu.L FLT3, 1. mu.M TK-S, 2. mu.M ATP; 2% DMSO, 0.4 ng/. mu.L FLT3-D835Y, 1. mu.M TK-S, 1. mu.M ATP; 2% DMSO, 0.2 ng/. mu. L c-KIT, 1. mu.M TK-S, 20. mu.M ATP. After incubation for 1 hour at room temperature, the antibody was added for detection. And simultaneously setting a solvent control group, a Gilteritinib positive control group and a blank control group which replace the compound to be detected with DMSO, and setting 3 multiple wells for each concentration of each sample. The activity of the sample is tested under a single concentration condition, e.g., 10. mu.M, selected for the primary screen. For samples that exhibit activity under certain conditions, e.g., an Inhibition% Inhibition greater than 50, the activity dose dependence, i.e., IC, is tested₅₀Values, obtained by nonlinear fitting of sample concentrations through sample activity, were calculated as Graphpad Prism 5, the model used for fitting was sigmoidal dose-response (variable slope), and for most inhibitor screening models, the bottom and top of the fitted curve were set at 0 and 100.

IC of Table 1 Compounds on FLT3, FLT3-D835Y, c-KIT kinase₅₀(nM)

Inhibition activity test for cell proliferation

MTS assay for in vitro proliferation inhibitory Activity (IC) of Compounds of the present invention against test cell lines₅₀): cells in logarithmic growth phase were trypsinized, counted and counted at 1X 10⁴Cell/well Density was seeded in 96-well plates, 100. mu.L per well, in a medium containing 5% CO₂Culture at 37 deg.CThe culture was carried out overnight in a incubator with six concentration gradients for each compound and three sets of multiple wells for each concentration, and after addition, the culture was carried out for 72 hours with 20. mu.L MTS added. After incubation for 2 hours at 37 ℃, the light absorption value at 490nm (L1) is measured by a SpectraMAX 340 microplate reader, the reference wavelength is 690nm (L2), the values (L1-L2) are plotted against different inhibitor concentrations, and the half inhibition concentration IC is obtained by formula fitting₅₀。

Table 2 IC of representative Compounds on acute myeloid leukemia cell MV4-11₅₀(nM)

TABLE 3 IC of representative Compounds on human leukemia cells Kasumi-1₅₀(nM)

Table 4 IC of representative Compounds on human mantle cell lymphoma cell Z138₅₀(nM)

IC of representative Compounds of Table 5 on human Breast cancer cells MCF-7₅₀(nM)

Preliminary pharmacodynamic results show that the compound has the effect of inhibiting FLT3, has certain proliferation inhibition activity on FLT3-D835Y mutation, has moderate inhibition strength on acute myelogenous leukemia cells, human leukemia cells Kasumi-1 and human mantle cell lymphoma cells Z138, and also has moderate inhibition strength on breast cancer MCF-7 cell strains, and can be used for preventing or treating clinical diseases related to FLT3 inhibition, wherein the diseases can be breast cancer, ovarian cancer, lung cancer, prostatic cancer, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, leukemia, lymphoma, neuroblastoma, glioma, head and neck cancer, thyroid cancer, liver cancer, cervical cancer, bladder cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumor, skin cancer and multiple myeloma.

Claims

1. A kind ofN ²-derivatives of carbamoyl aromatic ring-2-aminopyrimidines, characterized by having the general formula II:

and optical isomers and pharmaceutically acceptable salts thereof,

R₁selected from NR_d(CH₂)_nR_c、

；

R_cselected from unsubstituted or substituted C_5-8Cycloalkyl, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkyl, unsubstituted or substituted C₈-₁₈Hetero atom-containing spirocycloalkyl group, unsubstituted or substituted C_6-12Bridged cycloalkyl in which the heteroatom is at least one of nitrogen and oxygen, said substituent being selected from C_1-6Alkyl, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_f；

m is an integer of 0;

R_dand R_fEach independently selected from H, deuterium;

R₂is selected from-CONR_aR_b、-SO₂NR_aR_b、-NR_aCOR_b、-NR_aSO₂R_b；

R_aAnd R_bEach independently selected from H, deuterium, C_1-6Alkyl, unsubstituted C_3-8An aliphatic ring group;

R₃and R₄Each independently selected from H, deuterium.

2.N ²-carbamoyl aromatic ring-2-aminopyrimidine derivatives, optical isomers and pharmaceutically acceptable salts thereof, characterized by being selected from:

3.N ²-derivatives of carbamoyl aryl-2-aminopyrimidines, characterized by having the general formula III:

and optical isomers and pharmaceutically acceptable salts thereof,

R₁is selected from NR_d(CH₂)_nR_c、

；

n is an integer of 0 or 3; h and k are each independently selected from integers of 1 to 4;

R_cselected from substituted C_5-8Cycloalkyl, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkyl, unsubstituted or substituted C₈-₁₈Hetero atom-containing spirocycloalkyl group, unsubstituted or substituted C_6-12Bridged cycloalkyl in which the heteroatom is at least one of nitrogen and oxygen, the substituent being selected from C_1-6Alkyl, halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_f；

m is an integer of 0;

R_dand R_fEach independently selected from H, deuterium;

R₂is selected from-CONR_aR_b、-SO₂NR_aR_b、-NR_aCOR_b、-NR_aSO₂R_b；

R₃and R₄Each independently selected from H, deuterium, C_1-6Alkyl radical, C_1-6An alkoxy group.

4.N ²-carbamoyl aromatic ring-2-aminopyrimidine derivatives, optical isomers and pharmaceutically acceptable salts thereof, characterized by being selected from:

5.N ²-derivatives of carbamoyl aromatic ring-2-aminopyrimidines, characterized by having the general formula IV:

and optical isomers and pharmaceutically acceptable salts thereof,

q is selected from six-membered cyclic lactams;

R₁selected from NR_d(CH₂)_nR_c;

n is an integer of 0;

R_cselected from substituted C_5-8Cycloalkyl, unsubstituted or substituted C_5-8Heterocycloalkyl, unsubstituted or substituted C_8-18Spirocycloalkyl, unsubstituted or substituted C₈-₁₈Hetero atom-containing spirocycloalkyl group, unsubstituted or substituted C_6-12Bridged cycloalkyl wherein the heteroatom is nitrogen, said substituent being selected from the group consisting of halogen, (CH)₂)_mOH、(CH₂)_mNR_dR_f；

m is an integer of 0;

R_dand R_fEach independently selected from H, deuterium;

R₃and R₄Each independently selected from H, deuterium, C_1-6Alkyl, halogenated C_1-6Alkyl radical, C_1-6Hydroxyalkyl radical, C_3-8Cycloalkane radical, via C_1-6Alkyl chain linked C_3-8Cycloalkyl, cyano.

6.N ²-carbamoyl aromatic ring-2-aminopyrimidine derivatives, optical isomers and pharmaceutically acceptable salts thereof, characterized by being selected from:

7.N ²-carbamoyl aromatic ring-2-aminopyrimidine derivatives, optical isomers and pharmaceutically acceptable salts thereof, characterized by being selected from:

。

8. the method according to any one of claims 1 to 7N ²The application of the carbamyl aromatic ring-2-aminopyrimidine derivatives in preparing antitumor drugs is characterized in that the derivatives comprise optical isomers and pharmaceutically acceptable salts thereof, wherein the salts are organic acid salts and inorganic acid salts, and the organic acid salts are selected from citrate, fumarate, oxalate, malate, lactate, camphorsulfonate, p-toluenesulfonate, methanesulfonate, benzoate, tartrate, succinate, maleate, formate, acetate and amino acid salts; wherein the inorganic acid salt is selected from halogen acid salt, sulfate, phosphate and nitrate.

9. Use according to claim 8, wherein the amino acid salt is selected from the group consisting of aspartate and hydrohalide salts are selected from the group consisting of hydrochloride.

10. Use according to claim 8, wherein the medicament is used alone and/or in combination with other chemotherapeutic agents, surgical therapy, radiotherapy and the tumour is leukaemia, lymphoma, breast cancer.